上海交通大学学报(医学版) ›› 2021, Vol. 41 ›› Issue (11): 1454-1460.doi: 10.3969/j.issn.1674-8115.2021.11.008

• 论著 · 基础研究 • 上一篇    下一篇

敲减免疫调节蛋白B7-H3基因对膀胱癌细胞恶性增殖、侵袭和干细胞样特性的影响

王晓寒1(), 叶云林2, 肖康华2, 赵志鑫3, 吴静雅4   

  1. 1.河南护理职业学院外科教研室,安阳 455000
    2.中山大学附属肿瘤医院泌尿肿瘤科,广州 510030
    3.河南护理职业学院药学教研室,安阳 455000
    4.河南省安阳地区医院中医科,安阳 455000
  • 出版日期:2021-11-28 发布日期:2021-12-03
  • 通讯作者: 王晓寒 E-mail:wang136473@163.com
  • 作者简介:王晓寒(1983—),女,副教授,硕士;电子信箱:wang136473@163.com
  • 基金资助:
    2018年度河南省医学科技攻关计划项目(2018020950)

Effects of knockdown of immunoregulatory protein B7-H3 gene on malignant proliferation, invasion and stem cell-like characteristics of bladder cancer cells

Xiao-han WANG1(), Yun-lin YE2, Kang-hua XIAO2, Zhi-xin ZHAO3, Jing-ya WU4   

  1. 1.Department of surgery, Henan Vocational College of nursing, Anyang 455000, China
    2.Department of Urology and oncology, Cancer Hospital Affiliated to Zhongshan University, Guangzhou 510030, China
    3.Pharmacy teaching and Research Office, Henan Vocational College of nursing, Anyang 455000, China
    4.Department of traditional Chinese medicine, Henan Province Anyang District Hospital, Anyang 455000, China
  • Online:2021-11-28 Published:2021-12-03
  • Contact: Xiao-han WANG E-mail:wang136473@163.com
  • Supported by:
    2018 Henan Province Medical Science and Technology Tackling Project(2018020950)

摘要:

目的·探讨干扰免疫调节蛋白B7-H3基因对膀胱癌J82细胞的恶性增殖和干细胞样特性的影响。方法·shRNA干扰效率的检测采用反转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)。J82细胞增殖采用克隆形成实验检测。细胞凋亡采用流式细胞术检测。细胞侵袭采用Transwell检测。肿瘤克隆能力采用肿瘤成球实验观察。细胞凋亡、侵袭和干细胞样相关蛋白表达采用蛋白质印迹法(Western blotting)检测。结果·B7-H3在膀胱癌组织及膀胱癌细胞系中高表达(P=0.000)。确定shRNA2为设计的3组shRNAs中干扰B7-H3基因效率最高(P=0.000)。B7-H3干扰抑制了J82细胞的增殖、侵袭、肿瘤克隆能力及干细胞样特性(P=0.000)。并且,B7-H3干扰促进了J82细胞的凋亡(P=0.000)。结论·免疫调节蛋白B7-H3基因干扰可对膀胱癌J82细胞的恶性增殖、侵袭及干细胞样特性产生抑制作用。

关键词: B7-H3, 膀胱癌, J82细胞, 干细胞样特性

Abstract:

Objective·To investigate the effect of gene interfering with immune regulatory protein B7-H3 on malignant proliferation and stemness characteristics of bladder cancer J82 cells.

Methods·Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect shRNA interference efficiency. The proliferation of J82 cells was detected by clone formation assay. Cell apoptosis was detected by flow cytometry. Cell invasion was detected using Transwell asssay. The tumor-cloning ability was observed using tumor sphere formation experiment. Apoptosis, invasion, and expression and stemness-related proteins were detected by Western blotting.

Results·B7-H3 is highly expressed in bladder cancer tissues and bladder cancer cell lines (P=0.000). shRNA2 was identified as the most efficient interference with B7-H3 gene in three shRNAs (P=0.000). The interference of B7-H3 inhibited the proliferation, invasion, tumor cloning ability and stem cell-like characteristics of J82 cells (P=0.000). Furthermore, B7-H3 interference promoted the apoptosis of J82 cells (P=0.000).

Conclusion·The gene interference of immunoregulatory protein B7-H3 can inhibit the malignant proliferation, invasion and stemness characteristics of bladder cancer J82 cells, providing a new possibility and effective evidence for the treatment of bladder cancer.

Key words: B7-H3, Bladder cancer, J82 cells, Stemness characteristics

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